Assay of ganglioside GM2-N-acetyl-beta-D-galactosaminidase activity in human fibroblasts employing the natural activator protein--diagnosis of variant forms of GM2 gangliosidosis.
The physiological activator protein for the degradation of ganglioside GM2 by hexosaminidase A has been employed to assess the capability of cultured human fibroblast extracts to catalyze this ganglioside. This method permits a more reliable diagnosis of the different variants of GM2 gangliosidoses than the methods hitherto used. These either rely on artificial substrates or, when natural substrates are used, on detergents. Our method avoids a number of possible sources of error introduced by the unphysiological detergents, such as alteration of the isoenzymes' substrate specificity or inactivation of the enzymes. The range of application of the new method is discussed.
D002478:Cells, Cultured; D055598:Chemical Phenomena; D002621:Chemistry; D004789:Enzyme Activation; D005347:Fibroblasts / Q000201:enzymology; D049289:G(M2) Activator Protein; D005678:G(M2) Ganglioside / Q000378:metabolism*; D005732:Gangliosides / Q000378:metabolism*; D005733:Gangliosidoses / Q000175:diagnosis / Q000201:enzymology*; D006023:Glycoproteins; D006596:Hexosaminidases / Q000032:analysis*; D006801:Humans; D008722:Methods; D011506:Proteins / Q000502:physiology*; D049231:Saposins; D049128:Sphingolipid Activator Proteins; D013657:Taurodeoxycholic Acid
D049289:G(M2) Activator Protein; D005732:Gangliosides; D006023:Glycoproteins; C483345:PSAP protein, human; D011506:Proteins; D049231:Saposins; D049128:Sphingolipid Activator Proteins; D005678:G(M2) Ganglioside; D013657:Taurodeoxycholic Acid; C028960:GM2-N-acetyl-beta-D-galactosaminidase; D006596:Hexosaminidases